Immortalizing cells.

Steven Loechel frosty at biobase.dk
Fri Mar 10 03:28:43 EST 1995


I'm about to start a new project that involves immortalizing primary
human cells (trophoblasts), and as a novice in this area of research,
I have some humble questions that I would like to pose.

1) Is transfection with a plasmid that expresses SV40 large T antigen
still the method of choice for immortalization of most primary cells?

2) These cells are apparently refractory to transfection by liposomes,
but electroporation seems to work pretty well for them. We're about to
buy an electroporator. Is the Biorad apparaturs as good as any?

3) These cells are extraordinarily sensitive to Geneticin. Is this a 
potential problem for selecting transformants carrying a neo-R 
plasmid? 

If these are questions that have already been addressed ad nauseum in
this newsgroup, please email me at frosty at biobase.dk, or direct me to the
FAQ for this newsgroup.

Thanks,
Frosty



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